重复性的动脉剥脱损伤腹主动脉阻断在小鼠模型
Summary
理解再内皮的细胞和分子机制下列动脉剥蚀损伤是至关重要的在防止血栓形成和动脉的再狭窄。在这里，我们描述了肾腹主动脉动脉可重复性损伤剥蚀的协议。该程序的开发研究，规范使用小鼠模型内皮细胞再生的基本机制。
Abstract
经皮血管介入均匀导致动脉剥脱伤了随后导致血栓形成和再狭窄。这些并发症可在伤口边缘内再内皮归因于损伤。然而，再内皮的细胞和分子机制仍有待确定。虽然一些动物模型动脉剥脱术后学习再内皮可用，很少在鼠标由于局限性手术来执行。这破坏的机会，以利用转基因小鼠系，并调查特定基因的至再内皮过程中的贡献。在这里，我们提出了一个一步一步的协议中使用外部钳位血管下腹腹主动脉动脉创造剥蚀损伤的高度重复性的小鼠模型。纤维蛋白原和β-catenin的受伤主动脉的免疫细胞化学染色表现出亲血栓表面的暴露ð内皮完整的边界，分别。这里介绍的方法具有速度，优良的整体存活率，和相关技术容易的优点，创造了在转基因小鼠模型强加动脉剥蚀损伤一个唯一实用工具。使用这种方法，调查人员可能阐明正常或病理状态下重新内皮化的机制。
Introduction
血栓形成和再狭窄的患者严重的早期和晚期并发症接受谁经皮血管介入，如血管内球囊成形术1,2。多种策略已经被用来解决这些并发症，尤其是双重抗血小板治疗和药物洗脱支架。然而，很少焦点一直放在血栓形成和再狭窄，内皮细胞覆盖率（剥蚀）即损失的潜在原因。剥蚀损伤是由于机械外伤到血管壁介入过程的必然结果。这种机械创伤可导致损坏和去除基底膜和血管平滑肌的保护内皮细胞层并且暴露于循环血液3。在这些领域中的内皮细胞的损失创建前血栓形成和促炎环境，不仅促进血小板粘附和随后的血栓形成，但LSO刺激迁移，并导致新内膜增厚和狭窄4血管平滑肌细胞的增殖。这些并发症，以及它们的相关的治疗，导致显著发病率，最值得注意的是复发性缺血性疾病和出血事件影响人类健康。
再内皮从伤口边缘的裸露损伤是非常重要的预防血栓形成和再狭窄5。尸检结果和动物模型已经证明有效的减少血栓的发生率与支架支柱覆盖6,7。药物洗脱支架，设计通过抑制平滑肌细胞增殖和内膜增生减少再狭窄率，导致动脉再内皮损伤显著和晚期血栓形成3率持续上升。不幸的是，对于认识再内皮化的机制一直是一个缓慢的过程，由于缺乏AP的主要限于propriate动物模型8。
几种动物模型对于理解下面的动脉损伤血管内皮细胞和血管平滑肌细胞的作用已创建7,9,10。大鼠颈动脉球囊损伤模型是最好的特点，并已用于研究在总量，细胞和分子水平11剥蚀损伤的作用。然而，具有优良的存活率动脉剥脱伤的高度重复性的小鼠模型的缺少和急需采取的提供给多个设置更好地阐明血管再生多个转基因株系的优势。
这份手稿介绍动脉剥脱伤是可重复的，简单执行的小鼠模型。该方法已经显示出最小的发病率和死亡率在几个转基因株系。因为转基因小鼠系的广泛数量的，该模型可以用于阐明剥蚀损伤后再内皮的分子机制。
Protocol
Representative Results
Discussion
动脉损伤剥蚀，由于经皮干预措施，例如气囊血管成形术和血管支架，导致早期和晚期血管栓塞和再狭窄，并有助于复发性缺血事件3,12。有趣的是，外科血管夹紧也被牵连作为动脉剥蚀的原因，扩大的问题，以进行任何血管过程的患者的范围内，无论是经皮或打开13。而受损的再内皮化是血栓形成和再狭窄的相当认可的起因，周围再内皮化的分子机制已难以澄清。动物模型已成为…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作是由从礼与再生医学的Edythe广泛中心补助和与健康（HL130290）的国家机构来支持MLIA加州大学洛杉矶分校训练计划干细胞研究，以ASS和AIM，菲利普·J.·惠特科姆奖学金到AIM。
Materials
| Zeiss Discovery.V12 Stereomicroscope | Zeiss | 495037-9904-000 | |
| Rodent Heated Surgical Platform | Protech International | RES4000 | Heated platform for body temperature maintenance with nosecone for anesthestic maintenance and on which surgical procedure is performed |
| Isoflurane | Henry Schein | 50033 | 4% Induction; 2.5% Maintenance |
| Isoflurane Vaporizer | Summit Medical Equipment | 470062 | |
| Stryker T/Pump Warm Water Recirculator | Kent Scientific | TP-700 | |
| Artifical Tears Lubricant Opthalmic Ointment | Akorn Animal Health | 17478-162-35 | |
| Carprieve (Carprofen) | Norbrook Laboratories | NDC 55529-131-01 | |
| Oster™ A5 Professional Animal Clipper | M.Schneider & Sons Inc. | 78005010 | Use with animal clipper size 40 |
| Adjustable Wire Retractor | Fine Science Tools | 17004-05 | |
| Schwartz Micro Serrefines – Sharp Bend | Fine Science Tools | 18052-03 | |
| Surgical Instruments | Fine Science Tools | sharp dissecting forceps, blunt forceps, fine scissors, spring scissors, hemostat | |
| 0.5% Marcaine | Hospira | 0409-1610-50 | |
| 5-0 Suture, Vicryl | Fisher Scientific | NC0189890 | tapered needle |
| Vetbond | Fisher Scientific | NC0304169 | |
| Falcon® 35 mm Not TC-Treated Easy-Grip Style Bacteriological Petri Dish | Corning | 351008 | |
| Sylgard 184 Silicone Elastomer Kit | Dow Corning | 3097358-1004 | |
| Dissecting pins | Fisher Scientific | NC9681411 | |
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